人食管癌Eca-109细胞膜蛋白分离纯化方法的研究

The study on isolation and purification of membrane proteins of human Eca-109 cell line

目的 建立人食管癌细胞株Eca 10 9细胞膜蛋白分离和初步纯化方法。方法 使用改良的Neville法提取细胞膜 ,在pH 6.3的条件下 ,用去垢剂TritonX 10 0和辛基 β 葡糖苷把细胞膜上的蛋白溶解下来 ,以超滤法纯化膜蛋白并分组。 结果 在pH6.3的条件下 ,适当的去垢剂与膜蛋白之比 (即mg去垢剂 /mg膜蛋白 ) ,使用辛基 β 葡糖苷时为 6∶1;使用TritonX 10 0时为 2∶1。超滤法将膜蛋白分为 <3KD、3～ 10KD、<10KD、10～ 3 0KD、3 0～ 10 0KD、>10 0KD共 6个组。结论 为进一步研究食管癌Eca 10

Objective To establish the methods used to isolate and purify the membrane proteins wtth different molecular weight in human esophageal carcinoma Eca-109 cell line.Methods The cell membrane was isolated by the modified Neville' s method (pH 6.3) . The membrane proteins were dissolved out from cytomembrane with the detergents of Triton X-100 and Octyl-β-D-glucopyranoside respectively. Results In the Octyl-β-D-glucopyranoside group, appropriate detergent :protein ratio(milligram detergent:milligram membrane protein) was 6∶1.In the Triton X-100 group the ratio was 2∶1. Conclusion This study provides foundation to investigate the tumor antigen of human Eca-109 cell line.